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1、棉花学报CottonScience2014袁26渊3冤院189~196基因沉默对棉花幼苗生长和内源激素含量的影响1,211111*王丽袁穆春袁张明才袁杜明伟袁田晓莉袁李召虎(1.植物生长调节剂教育部工程研究中心/中国农业大学农学与生物技术学院袁北京100193曰2.河南师范大学生命科学学院袁河南新乡453007)摘要院利用RT-PCR从棉花中获得493bp的渊赤霉素渊GA冤合成酶基因冤片段袁经RⅠ/I双酶切后连入pTRV-RNA2质粒袁获得了重组载体pTRV-GhCPS曰用该载体转化农杆菌后袁室内盆栽条件下侵染棉花品种欣抗4的子叶苗遥结果表明袁病毒
2、诱导沉默基因的棉花幼苗中的表达量仅为空载体对照的52%遥与空载体对照相比袁沉默的棉花幼苗第1叶和第2叶的内源GA4含量分别降低33%和60%曰第2叶ABA含量降低48%袁IAA尧ZR和iPA含量均无显著变化曰株高降低64%遥第1叶和第2叶的叶面积分别减小26%和47%袁总叶绿素含量分别增加29%和63%袁类胡萝卜素含量分别增加27%和46%遥第1叶的净光合速率尧气孔导度尧胞间CO2和蒸腾速率分别增加23%尧40%尧9%和31%遥可见袁利用病毒诱导基因沉默技术技术沉默后可调控棉花幼苗叶片内源激素平衡袁控制棉花幼苗的生长袁并提高单位叶面积的光合能力遥关
3、键词院棉花曰病毒诱导的基因沉默曰赤霉素曰净光合速率曰ABA中图分类号院S562.01文献标志码院A文章编号院1002-7807(2014)03-0189-081,211111*WangLi,MuChun,ZhangMingcai,DuMingwei,TianXiaoli,LiZhaohu(1.100193,;2.453007,)Virus-inducedgenesilencing(VIGS)isanewreversegeneticsmethodusedinrapididentificationofgenefunctioninplantandhasb
4、ecomeoneofthemostwidelyusedplantfunctionalgenomicstools.Ent-copalyldiphosphatesynthase(CPS)isthefirstkeyenzymeinvolvedingibberellin(GA)biosynthesis.WeaimedtoelucidatetheeffectofsilencingbyVIGSonthegrowthandendogenoushormonecontentofcottonseedlings.Inthisstudy,a493-bpfragmentof
5、wasamplifiedfromthecottoncultivar(Xinkang4)cDNAusingRT-PCRandwassubsequentlyinsertedintothepTRV-RNA2vectorusingRⅠandIsites,respectively.pTRV-GhCPSconstructwastransformedintoGV3101andwasusedtotransformcottonseedlingsatthecotyledonstagewithcottoncultivarXinkang4growninagrowthcha
6、mberunderpotculture.TheresultsshowedthattheexpressionlevelofinVIGSplantswasonly52%ofthatinpTRVcontrolplants.Si-lencingresultedin33%and60%reductionsinGA4contentinthe1stand2ndleaves,respectively,andaslightdecreaseofIAAcontentbutnoobviouschangesinZRandiPAcontentsinbothleaves.ABAc
7、ontentinthe2ndleafwasdecreasedby48%.TheheightofVIGSplantswasreducedby64%.Theareaofthe1stand2ndleavesinVIGSplantswasde-creasedby26%and47%,respectively.Silencingincreasedtotalchlorophyllcontentsby29%and63%,andcarotenoidscontentsby27%and46%inthe1stand2ndleaves,respectively.Netpho
8、tosynthesisrate,stomatalconductance,intercellularCO2concentra